Spatio-Temporal distribution of a vector of cutaneous leishmaniasis: Pintomyia longiflocosa, in a population from the Colombian Andean Mountains.

BACKGROUND: Leishmaniasis, a neglected disease and public health concern, is associated with various factors such as biological, social, economical conditions and climate, increasing the risk of human infection. Understanding the population dynamics of the vectors, like Pintomyia longiflocosa, and its relationship with ecological variables is crucial for developing effective strategies to control sand fly populations and combat cutaneous leishmaniasis in a tropical country like Colombia. METHODOLOGY: Adult sand flies were collected in three different sample locations: outdoor, indoor, and peri-domestic areas in three houses located in the rural settlement of Campoalegre (Huila) between February 2020 and February 2021, using the CDC light traps. The sand fly density was quantified and associated with the sample locations and the sampling months using Analysis of Variance and Pearson correlations. PRINCIPAL FINDINGS: In the period of the sample, 98.86% of sand fly collected was identified as Pi. longiflocosa. The density of this species was significantly different between males and females, the latter contributing more to density in all sample locations (P<0.0001). The outdoor was the sample location with the highest and most significative density in this study (70%, P = 0.04). The density of these sand flies is related to the seasonality of Campoalegre, revealing a density peak from February and June to October (P < 0.05). Finally, precipitation is the environmental variable prominently linked to the density pattern, showing a negative correlation with it. Months with the highest precipitations show the lowest values of Pi. longiflocosa abundance. CONCLUSIONS/SIGNICANCE: Our investigation reveals a inverse correlation between precipitation levels and the abundance of Pi. longiflocosa in Campoalegre (Huila), particularly in outdoor areas. This suggests that vector control strategies to periods of reduced precipitation in outdoor settings could offer an effective approach to minimizing cases of cutaneous leishmaniasis in the region.

High Levels of Diversity in Anopheles Subgenus Kerteszia Revealed by Species Delimitation Analyses.

The Anopheles subgenus Kerteszia is a poorly understood group of mosquitoes that includes several species of medical importance. Although there are currently twelve recognized species in the subgenus, previous studies have shown that this is likely to be an underestimate of species diversity. Here, we undertake a baseline study of species delimitation using the barcode region of the mtDNA COI gene to explore species diversity among a geographically and taxonomically diverse range of Kerteszia specimens. Beginning with 10 of 12 morphologically identified Kerteszia species spanning eight countries, species delimitation analyses indicated a high degree of cryptic diversity. Overall, our analyses found support for at least 28 species clusters within the subgenus Kerteszia. The most diverse taxon was Anopheles neivai, a known malaria vector, with eight species clusters. Five other species taxa showed strong signatures of species complex structure, among them Anopheles bellator, which is also considered a malaria vector. There was some evidence for species structure within An. homunculus, although the results were equivocal across delimitation analyses. The current study, therefore, suggests that species diversity within the subgenus Kerteszia has been grossly underestimated. Further work will be required to build on this molecular characterization of species diversity and will rely on genomic level approaches and additional morphological data to test these species hypotheses.

Morphology of the larvae, male genitalia and DNA sequences of Anopheles (Kerteszia) pholidotus (Diptera: Culicidae) from Colombia.

Since 1984, Anopheles (Kerteszia) lepidotus has been considered a mosquito species that is involved in the transmission of malaria in Colombia, after having been incriminated as such with epidemiological evidence from a malaria outbreak in Cunday-Villarrica, Tolima. Subsequent morphological analyses of females captured in the same place and at the time of the outbreak showed that the species responsible for the transmission was not An. lepidotus, but rather Anopheles pholidotus. However, the associated morphological stages and DNA sequences of An. pholidotus from the foci of Cunday-Villarrica had not been analysed. Using samples that were caught recently from the outbreak region, the purpose of this study was to provide updated and additional information by analysing the morphology of female mosquitoes, the genitalia of male mosquitoes and fourth instar larvae of An. pholidotus, which was confirmed with DNA sequences of cytochrome oxidase I and rDNA internal transcribed spacer. A total of 1,596 adult females were collected in addition to 37 larval collections in bromeliads. Furthermore, 141 adult females, which were captured from the same area in the years 1981-1982, were analysed morphologically. Ninety-five DNA sequences were analysed for this study. Morphological and molecular analyses showed that the species present in this region corresponds to An. pholidotus. Given the absence of An. lepidotus, even in recent years, we consider that the species of mosquitoes that was previously incriminated as the malaria vector during the outbreak was indeed An. pholidotus, thus ending the controversy.

Morphology of the larvae, male genitalia and DNA sequences of Anopheles (Kerteszia) pholidotus (Diptera: Culicidae) from Colombia

Since 1984, Anopheles (Kerteszia) lepidotus has been considered a mosquito species that is involved in the transmission of malaria in Colombia, after having been incriminated as such with epidemiological evidence from a malaria outbreak in Cunday-Villarrica, Tolima. Subsequent morphological analyses of females captured in the same place and at the time of the outbreak showed that the species responsible for the transmission was not An. lepidotus, but rather Anopheles pholidotus. However, the associated morphological stages and DNA sequences of An. pholidotus from the foci of Cunday-Villarrica had not been analysed. Using samples that were caught recently from the outbreak region, the purpose of this study was to provide updated and additional information by analysing the morphology of female mosquitoes, the genitalia of male mosquitoes and fourth instar larvae of An. pholidotus, which was confirmed with DNA sequences of cytochrome oxidase I and rDNA internal transcribed spacer. A total of 1,596 adult females were collected in addition to 37 larval collections in bromeliads. Furthermore, 141 adult females, which were captured from the same area in the years 1981-1982, were analysed morphologically. Ninety-five DNA sequences were analysed for this study. Morphological and molecular analyses showed that the species present in this region corresponds to An. pholidotus. Given the absence of An. lepidotus, even in recent years, we consider that the species of mosquitoes that was previously incriminated as the malaria vector during the outbreak was indeed An. pholidotus, thus ending the controversy.

Anthropophilic biting behaviour of Anopheles (Kerteszia) neivai Howard, Dyar & Knab associated with Fishermen’s activities in a malaria-endemic area in the Colombian Pacific

On the southwest Pacific Coast of Colombia, a field study was initiated to determine the human-vector association between Anopheles (Kerteszia) neivai and fishermen, including their nearby houses. Mosquitoes were collected over 24-h periods from mangrove swamps, marshlands and fishing vessels in three locations, as well as in and around the houses of fishermen. A total of 6,382 mosquitoes were collected. An. neivai was most abundant in mangroves and fishing canoes (90.8%), while Anopheles albimanus was found indoors (82%) and outdoors (73%). One An. neivai and one An. albimanus collected during fishing activities in canoes were positive for Plasmodium vivax , whereas one female An. neivai collected in a mangrove was positive for P. vivax . In the mangroves and fishing canoes, An. neivai demonstrated biting activity throughout the day, peaking between 06:00 pm-07:00 pm and there were two minor peaks at dusk and dawn. These peaks coincided with fishing activities in the marshlands and mangroves, a situation that places the fishermen at risk of contracting malaria when they are performing their daily activities. It is recommended that protective measures be implemented to reduce the risk that fishermen will contract malaria.

Anthropophilic biting behaviour of Anopheles (Kerteszia) neivai Howard, Dyar & Knab associated with Fishermen's activities in a malaria-endemic area in the Colombian Pacific.

On the southwest Pacific Coast of Colombia, a field study was initiated to determine the human-vector association between Anopheles (Kerteszia) neivai and fishermen, including their nearby houses. Mosquitoes were collected over 24-h periods from mangrove swamps, marshlands and fishing vessels in three locations, as well as in and around the houses of fishermen. A total of 6,382 mosquitoes were collected. An. neivai was most abundant in mangroves and fishing canoes (90.8%), while Anopheles albimanus was found indoors (82%) and outdoors (73%). One An. neivai and one An. albimanus collected during fishing activities in canoes were positive for Plasmodium vivax , whereas one female An. neivai collected in a mangrove was positive for P. vivax . In the mangroves and fishing canoes, An. neivai demonstrated biting activity throughout the day, peaking between 06:00 pm-07:00 pm and there were two minor peaks at dusk and dawn. These peaks coincided with fishing activities in the marshlands and mangroves, a situation that places the fishermen at risk of contracting malaria when they are performing their daily activities. It is recommended that protective measures be implemented to reduce the risk that fishermen will contract malaria.

[Characteristics of new cell cultures derived from embryonic tissues of Aedes aegypti (Diptera: Culicidae)]. / Características de nuevos cultivos celulares derivados de tejidos embrionarios de Aedes aegypti (Diptera: Culicidae).

INTRODUCTION: Cell cultures from insects are a useful methodology in technological and biomedical studies. OBJECTIVE: The present work was aimed at obtaining and characterizing cell cultures derived from Aedes aegypti embryonic tissues. MATERIALS AND METHODS: Embryonated eggs were used for embryonic tissue explants in L-15/Grace and MMNP12 culture media, supplemented with 20% fetal bovine serum and a mixture of 1% antimycotic and antibiotics, at a pH ranging from 6.8 to 7.0. The incubation temperature was 28 degrees C; a CO2 atmosphere was not required. RESULTS: Cell growth was obtained in L-15/Grace medium three weeks after embryonic tissues explants. Six months were required for achieving a confluent monolayer. Twenty-eight serial cell subcultures were carried out from August 2003 to June 2004. Cell morphology was characterized as epithelial in the later subcultures. Karyotype morphometry as well as molecular and isozymatic profiles were established. The cultures were compared with adult samples from the species taken from the same colony and with cell lines derived from other insects. DISCUSSION AND CONCLUSIONS: These cells are an important in vitro system in applied and basic research.

Colonization of Ochlerotatus taeniorhynchus from Riohacha, Colombia.

The main objective of the present work was to establish and maintain a colony of Ochlerotatus taeniorhynchus, Riohacha strain, under laboratory conditions and to study its life cycle. The colony's insectary was established from adult male and female mosquitoes collected from peripheral neighborhoods of Riohacha, Colombia. Environmental conditions established in the insectary were 26.5 degrees C average temperature, 80% average relative humidity, and 12 h photoperiodicity. Eight continuous generations were taken into account for maintaining the mosquitoes and analyzing their life cycle. The male mosquito's average life cycle was 26.8 days. The female's cycle was 30.8 days. Analysis of each of the biological stages of development (mean days) produced the following results: egg incubation 4.55 +/- 0.291, larvae 8.28 +/- 0.499, pupae 1.32 +/- 0.215, adult male 12.65 +/- 5.920. and adult female 16.73 +/- 6.034. The Riohacha colony has been maintained for 32 generations in 31 months. Comparison of the Riohacha colony with the previously established Cartagena and Barranquilla colonies showed few differences in the duration of stages of the life cycle between strains.

Características de nuevos cultivos celulares derivados de tejidos embrionarios de Aedes aegypti (Diptera: Culicidae) / Characteristics of new cell cultures derived from embryonic tissues of Aedes aegypti (Diptera: Culicidae)

Introducción. Los cultivos celulares de insectos son una metodología útil en estudios biomédicos y tecnológicos.Objetivo. El propósito principal del presente trabajo fue obtener y caracterizar cultivos celulares derivados de tejidos embrionarios de Aedes aegypti. Materiales y métodos. Se emplearon huevos embrionados para los explantes de tejidos en los medios de cultivos MM/VP12 y L-15/Grace, con suplemento de 20 por ciento de suero fetal bovino y una mezcla al 1 por ciento de antibiótico y antimicótico, con un rango de pH entre 6,8 y 7,0. Los cultivos se incubaron a una temperatura de 28oC sin atmósfera de CO2. Resultados. El crecimiento celular se obtuvo en el medio L-15/Grace, 3 semanas después de haber sido sembrados los tejidos embrionarios; sin embargo, se necesitaron 6 meses para la formación de la monocapa confluente. Desde agosto de 2003 hasta junio de 2004, se habían realizado 28 subcultivos. Las células se caracterizaron morfológicamente; predominaron las formas epitelioides en subcultivos de pases altos. También se reconocieron las particularidades morfométricas del cariotipo y, además, se determinaron los perfiles isoenzimáticos y moleculares de los cultivos celulares, los cuales se compararon con muestras de adultos de la especie tomadas de la misma colonia y con líneas celulares derivadas de otros insectos.Discusión y conclusiones. Estas células representan, potencialmente, un importante sistema in vitro en investigaciones básicas y aplicadas

Life tables and reproductive parameters of Lutzomyia spinicrassa (Diptera: Psychodidae) under laboratory conditions.

Lutzomyia spinicrassa is a vector of Leishmania braziliensis in Colombia. This sand fly has a broad geographical distribution in Colombia and Venezuela and it is found mainly in coffee plantations. Baseline biological growth data of L. spinicrassa were obtained under experimental laboratory conditions. The development time from egg to adult ranged from 59 to 121 days, with 12.74 weeks in average. Based on cohorts of 100 females, horizontal life table was constructed. The following predictive parameters were obtained: net rate of reproduction (8.4 females per cohort female), generation time (12.74 weeks), intrinsic rate of population increase (0.17), and finite rate of population increment (1.18). The reproductive value for each class age of the cohort females was calculated. Vertical life tables were elaborated and mortality was described for the generation obtained of the field cohort. In addition, for two successive generations, additive variance and heritability for fecundity were estimated.

Life tables and reproductive parameters of Lutzomyia spinicrassa (Diptera: Psychodidae) under laboratory conditions

Lutzomyia spinicrassa is a vector of Leishmania braziliensis in Colombia. This sand fly has a broad geographical distribution in Colombia and Venezuela and it is found mainly in coffee plantations. Baseline biological growth data of L. spinicrassa were obtained under experimental laboratory conditions. The development time from egg to adult ranged from 59 to 121 days, with 12.74 weeks in average. Based on cohorts of 100 females, horizontal life table was constructed. The following predictive parameters were obtained: net rate of reproduction (8.4 females per cohort female), generation time (12.74 weeks), intrinsic rate of population increase (0.17), and finite rate of population increment (1.18). The reproductive value for each class age of the cohort females was calculated. Vertical life tables were elaborated and mortality was described for the generation obtained of the field cohort. In addition, for two successive generations, additive variance and heritability for fecundity were estimated.

Comparación cariotípica de cinco especies de Lutzomyia (Diptera: Psychodidae) de la serie townsendi grupo verrucarum en Colombia / Kariotypic comparison of five species of Lutzomyia (Diptera: Psychodidae) of the series townsendi and the verrucarum group in Colombia

Con el propósito de establecer algunas características citogenéticas de cinco especies nativas de Lutzomyia, correspondientes a la serie townsendi del grupo verrucarum: Lutzomyia longiflocosa, Lutzomyia quasitownsendi, Lutzomyia spinicrassa, Lutzomyia torvida y Lutzomyia youngi, se llevó a cabo un estudio comparativo entre los cariotipos y su morfometría cromosómica. A partir de ganglios cerebrales de larvas de IV estadio, se prepararon los cromosomas mitóticos mediante la técnica de aplastamiento (squash) del tejido. Se efectuaron las mediciones cromosómicas, atendiendo los siguientes par metros: brazo corto, brazo largo, relación de brazos, longitud total, longitud relativa, índice centromérico y longitud relativa promedio. Se clasificaron los cromosomas según su morfometría y posición del centrómero siguiendo patrones estandarizados. Se calculó la distancia taxonómica y con base en estos datos se separaron las especies y se ubicaron en un fenograma. Las cinco especies de flebótomos presentaron 4 pares de cromosomas, número diploide (2N8), y el número fundamental fue de 16. En ninguno de los cariotipos se observó heteromorfismo sexual cromosómico. El análisis estadístico de los datos de morfometría cromosómica mostró diferencias altamente significativas entre los pares cromosómicos de las cinco especies. Sin embargo, la longitud total del genoma en los flebotómos fue muy similar, a excepción de L. youngi. En conclusión, las especies íntimamente relacionadas se lograron diferenciar a nivel citotaxonómico.

Prevalencia de leishmaniasis visceral canina en municipios de Huila - Colombia / Prevalence of canine visceral leishmaniasis in municipalities of Huila, Colombia

Con el propósito de establecer la prevalencia de leishmaniasis canina se efectuó un estudio descriptivo en 307 caninos, ubicados en 17 veredas de los municipios de Neiva, Tello y Algeciras, a los cuales se les practicó examen clínico, punción y aspiración del ganglio linfático poplíteo para extendido y coloración en lámina; toma de sangre completa por venopunción para análisis mediante la técnica de inmunofluorescencia indirecta. La población canina estuvo conformada por perros mestizos con edad promedio de tres años: el 30,6/100 fueron hembras y el 69,4/100 machos. En el examen físico de los caninos se observó enflaquecimiento, 31/100; onicogrifosis, 29.3/100; linfadenitis de ganglio popliteo 3,2/100; areas alopécicas 21,8/100; lesiones eritematosas 11,5/100 y úlceras cutáneas 3,3/100. El 1,4/100 de los caninos analizados presentaron amastigotes en el aspirado y el 17,2/100 fueron seropositivos. La prevalencia de anticuerpos contra L.chagasi es alta comparada con otros estudios realizados en colombia y otros paises, corroborando la endemicidad de la enfermedad en el área. Por consiguiente, es necesario fortalecer las estrategias de vigilancia epidemiológica y la búsqueda activa de casos humanos.

[Karyotypic comparison of five species of Lutzomyia (diptera: psychodidae) of the series townsendi and the verrucarum group in Colombia]. / Comparación cariotípica de cinco especies de Lutzomyia (diptera: psychodidae) de la serie townsendi grupo verrucarum en Colombia.

Cytogenetic characteristics were established for five native species of phlebotomine sand flies (Lutzomyia, series townsendi, verrucarum group): Lutzomyia longiflocosa, Lutzomyia townsendi, Lutzomyia spinicrassa, Lutzomyia torvida and Lutzomyia youngi. Karyotypes and chromosomal morphometry were compared. Using the squash technique, brain tissues from late 4th instar larvae provided the necessary mitotic chromosomes. Chromosomal measurements were made on the following chromosomal characteristics: short arm, long arm, arm ratio, total length, relative length, centromeric index and relative length average of chromosomes. Chromosomes were classified according to their morphometry and position of the centromere. The taxonomic distance was calculated, and the relationships among the species displayed in a phenogram. All five species possessed four pairs of chromosomes as diploid number (2N = 8). None of the karyotypes indicated presence of heteromorphic chromosomes. Statistical analysis of the morphometric data showed highly significant differences among the chromosomes pairs of the five species. However, the total length of the genome was very similar, with the exception of L. youngi. In conclusion, these closely related species were distinguishable at cytological level.

A new continuous cell line from the mosquito Psorophora confinnis (Diptera: Culicidae) and its susceptibility to infections with some arboviruses

A new cell line, PC-0199-BR, was established from embryonated eggs of the mosquito Psorophora confinnis. To date (September 2000) it has had 62 continuous passages. This is the first report of a cell line of mosquitoes belonging to the genus Psorophora. Cell growth initially was achieved in the MM/VP12 medium, supplemented with 20 percent fetal bovine serum; however, the subcultures were later adapted to Grace's medium with 10 percent fetal bovine serum. Cell morphology in the primary cultures was heterogeneous; but later in the established cell line, the predominant cell type was epithelioid. Cultured cells were predominantly diploid (2n=6); however, chromosome abnormalities were observed in a small proportion of the cells in later passages. C and G band patterns were also determined in the karyotype. The cell line isozyme profiles coincided with pupae and adult samples of the species taken from the same colony. A preliminary arbovirus susceptibility study for the cell line was undertaken. No evidence was observed of contamination of the cell line with bacteria, fungi or mycoplasma